Optic glomeruli and their inputs in Drosophila share an organizational ground pattern with the antennal lobes

Studying the insect visual system provides important data on the basic neural mechanisms underlying visual processing. As in vertebrates, the first step in visual processing in insects is through a series of retinotopic neurons. Recent studies on flies have found that these converge onto assemblies of columnar neurons in the lobula, the axons of which segregate to project to discrete optic glomeruli in the lateral protocerebrum. This arrangement is much like the fly's olfactory system, in which afferents target uniquely identifiable olfactory glomeruli. Here, whole-cell patch recordings show that even though visual primitives are unreliably encoded by single lobula output neurons because of high synaptic noise, they are reliably encoded by the ensemble of outputs. At a glomerulus, local interneurons reliably code visual primitives, as do projection neurons conveying information centrally from the glomerulus. These observations demonstrate that in Drosophila, as in other dipterans, optic glomeruli are involved in further reconstructing the fly's visual world. Optic glomeruli and antennal lobe glomeruli share the same ancestral anatomical and functional ground pattern, enabling reliable responses to be extracted from converging sensory inputs.     

A new ascending sensory tract to the calyces of the honeybee mushroom body,the subesophageal-calycal tract

The mushroom bodies of the honeybee are important neuropils for learning and memory. Therefore, knowledge about their input and output connections is essential to understanding how these neuropils function. A newly described input tract to the mushroom body is presented here, which is called the subesophageal-calycal tract (SCT) and connects the subesophageal ganglion with the calyces of the mushroom bodies. The neuronal somata of the SCT neurons lie in one cluster between the lobula of the optic lobe and a neuropil area that is formed from the fusion of the tritocerebrum and the subesophageal ganglion. Within the subesophageal ganglion, the dendritic fibers of SCT neurons overlap with terminals of sensory neurons from the proboscis. Therefore, we conclude that the SCT neurons might process gustatory and mechanosensory information from the proboscis. Individual SCT neurons receive unilateral input within the subesophageal ganglion and may connect to either the ipsilateral or the contralateral mushroom body. On their way to the mushroom bodies, the SCT neuron axons meet the roots of the antennocerebralis tracts (ACTs) and from this point follow the same path as the median ACT neurons for a short distance. Within the calyces, the SCT neurons innervate two separate areas, a small area within the dorsal collar just below the lip and a part of the basal ring. Double-labeling experiments show that the projections of the SCT neurons do not overlap with the projections of the olfactory projection neurons and visual projection neurons from the dorsal medulla. The possible function of the SCT neurons and the relation of the SCT to known input tracts of the mushroom bodies in other insects are discussed.     

Serotonin-immunoreactive neurons in the brain of the honeybee

The distribution of serotonin-immunoreactive neurons in the brain of the worker honey bee Apis mellifera was studied by means of immunocytochemical staining by using a well-characterized antibody to serotonin (5-HT). About 75 immunoreactive perikarya are grouped into clusters in the optic lobe and in the median and dorsal protocerebrum. Immunoreactive fibers were resolved in all areas of the brain. The optic lobe shows restricted layers of 5-HT-immunoreactive fibers in the lamina and medulla organized perpendicular to the retinotopic elements. Immunoreactive fibers in the lobula represent invasions of protocerebral giant wide-field neurons. The nonglomerular neuropil of the brain exhibits a meshwork of immunoreactive fibres invading glomerular neuropil of the mushroom bodies, central body complex, and antennal lobes. Mushroom body stalks and lobes contain immunoreactive fibers arranged perpendicular to the Kenyon cell fibers and matching subcompartments of these corpora pedunculata areas. The calyces are devoid of immunofluorescence. Serotonin-positive fibres in the central body complex are arranged in its subcompartments. No 5-HT immunoreactivity was found in the pons. Antennal glomeruli contain immunoreactive fibers restricted around the margin of the glomeruli. The selective mapping of 5-HT-immunoreactive neurons complements studies on the distribution of monoamine-containing neurons in the bee brain. Serotonin- and catecholamine-containing neurons often occur together in the same brain areas and subcompartments. The immunohistochemical approach in chemoneuroanatomy gives new evidence for a more complicated architecture of the brain than could be deduced from the classical neuroanatomical studies.     

Structure and response patterns of olfactory interneurons in the honeybee, Apis mellifera

To analyze morphologic and physiological properties of olfactory interneurons in the honeybee, Apis mellifera, antennal lobe (AL) neurons were intracellularly recorded and subsequently labeled with Neurobiotin. Additional focal injections were carried out with cobalt hexamine chloride and dextran fluorescent markers. Olfactory interneurons (projection neurons, PNs) project by means of five tracts, the lateral, the median, and three mediolateral antennocerebral tracts (l-, m-, and ml-ACT, respectively) to the mushroom bodies (MBs) and the protocerebral lobe (PL) of the ipsilateral protocerebrum. Uniglomerular PNs of the m- and l-ACT receiving input from a single glomerulus of the AL also arborize in different regions of the AL. The vast majority of l-ACT innervate the T1 region, whereas m-ACT neurons arborize exclusively in the T2, T3, and T4 regions (T1-4 : AL projection area of sensory cells from the antennae). In the calyces of the MB, uniglomerular PNs form varicosities in the basal ring and the lip region. Individual neurons of both types exhibit unequal innervation within and between the two calyces. In addition, m-ACT fibers ramify more densely within the lip neuropil and show a higher incidence of spine-like processes than l-ACTs. In the PL, l-ACTs arborize exclusively within the lateral horn, whereas some m-ACT neurons innervate a broader region. Multiglomerular neurons of the ml-ACT leave the AL by means of three subtracts (ml-ACT 1-3). Two different types can be distinguished according to their protocerebral target areas: ml-ACTs projecting to the lateral PL (LPL) and to the neuropil around the alpha-lobe (tracts 2 and 3) and neurons projecting only to the LPL (tract 1). Intracellular recordings indicate that both l- and m-ACT neurons respond to general odors but with different response properties, indicating that odor information is processed in parallel pathways with different functional characteristics. Just like m-ACT neurons, ml-ACT neurons respond to odors with complex activity patterns. Bilateral interneurons, originating in the suboesophageal ganglion, connect glomeruli of both AL, and send an axon through the m-ACT in each hemisphere of the brain, terminating in the lip region of the calyces. These neurons respond to contact chemical stimuli.     

Systematic analysis of the visual projection neurons of Drosophila melanogaster. I. Lobula-specific pathways

In insects, visual information is processed in the optic lobe and conveyed to the central brain. Although neural circuits within the optic lobe have been studied extensively, relatively little is known about the connection between the optic lobe and the central brain. To understand how visual information is read by the neurons of the central brain, and what kind of centrifugal neurons send the control signal from the central brain to the optic lobe, we performed a systematic analysis of the visual projection neurons that connect the optic lobe and the central brain of Drosophila melanogaster. By screening approximately 4,000 GAL4 enhancer-trap strains we identified 44 pathways. The overall morphology and the direction of information of each pathway were investigated by expressing cytoplasmic and presynapsis-targeted fluorescent reporters. A canonical nomenclature system was introduced to describe the area of projection in the central brain. As the first part of a series of articles, we here describe 14 visual projection neurons arising specifically from the lobula. Eight pathways form columnar arborization in the lobula, whereas the remaining six form tangential or tree-like arborization. Eleven are centripetal pathways, among which nine terminate in the ventrolateral protocerebrum. Terminals of each columnar pathway form glomerulus-like structures in different areas of the ventrolateral protocerebrum. The posterior lateral protocerebrum and the optic tubercle were each contributed by a single centripetal pathway. Another pathway connects the lobula on each side of the brain. Two centrifugal pathways convey signals from the posterior lateral protocerebrum to the lobula.     

Immunocytochemistry of dopamine in the brain of the locust Schistocerca gregaria.

Catecholamine-induced histofluorescence studies have suggested a rich innervation of the locust brain by dopamine-containing neurons. To provide a basis for future studies on dopamine action in this insect, the location and morphology of neurons reacting with antisera against dopamine were investigated in the supraoesophageal ganglion of the locust, Schistocerca gregaria. In each brain hemisphere, about 100 interneurons in the midbrain and approximately 3,000 cells in the optic lobe show dopamine-like immunoreactivity. All major areas of the brain except the calyces of the mushroom body, the antennal lobe, large parts of the lobula, and some areas in the inferior lateral protocerebrum contain immunoreactive neuronal processes. The arborization patterns of most dopamine-immunoreactive cell types could be identified through detailed reconstructions. The central body exhibits the most intense immunostaining. It is innervated by at least 40 pairs of dopamine-immunoreactive neurons belonging to three different cell types. Additional arborizations of these neurons are in the superior protocerebrum and in the lateral accessory lobes. A group of 4 immunoreactive neurons with ramifications in the antennal mechanosensory and motor center gives rise to a dense meshwork of varicose fibers in the pedunculus and parts of the alpha- and beta-lobes of the mushroom body. Other cell types innervate the ventrolateral protocerebrum, the inferior protocerebrum and the posterior optic tubercles. Three descending neurons originating in the tritocerebrum exhibit dopamine-like immunoreactivity. In the optic lobe, about 3,000 columnar intrinsic neurons of the medulla and a group of centrifugal tangential cells with arborizations in the medulla and lamina are dopamine-immunoreactive.(ABSTRACT TRUNCATED AT 250 WORDS)     

Dual olfactory pathway in the honeybee, Apis mellifera

The antennal lobes (ALs) are the primary olfactory centers in the insect brain. In the AL of the honeybee, olfactory glomeruli receive input via four antennal sensory tracts (T1-4). Axons of projection neurons (PNs) leave the AL via several antenno-cerebral tracts (ACTs). To assign the input-output connectivity of all glomeruli, we investigated the spatial relationship of the antennal tracts and two prominent AL output tracts (medial and lateral ACT) mainly formed by uniglomerular (u) PNs using fluorescent tracing, confocal microscopy, and 3D analyses. Furthermore, we investigated the projections of all ACTs in higher olfactory centers, the mushroom-bodies (MB) and lateral horn (LH). The results revealed a clear segregation of glomeruli into two AL hemispheres specifically supplied by PNs of the medial and lateral ACT. PNs of the lateral ACT innervate glomeruli in the ventral-rostral AL and primarily receive input from T1 (plus a few glomeruli from T2 and T3). PNs of the medial ACT innervate glomeruli in the dorsal-caudal hemisphere, and mainly receive input from T3 (plus a few glomeruli from T2 and T4). The PNs of the m- and l-ACT terminate in different areas of the MB calyx and LH and remain largely segregated. Tracing of three mediolateral (ml) ACTs mainly formed by multiglomerular PNs revealed terminals in distinct compartments of the LH and in three olfactory foci within the lateral protocerebrum. The results indicate that olfactory input in the honeybee is processed via two separate, mainly uPN pathways to the MB calyx and LH and several pathways to the lateral protocerebrum.     

Dye fills reveal additional olfactory tracts in the protocerebrum of wild‐type Drosophila

The antennal lobe (AL) is the primary olfactory center in insect brains. It receives sensory input from the olfactory sensory neurons (OSNs) and sends, through its projection neurons (PNs), reformatted output to secondary olfactory centers, including the mushroom body (MB) calyx and the lateral horn (LH) in the protocerebrum. By injecting dye into the AL of wild‐type Drosophila, we identified previously unknown direct pathways between the AL and the ventrolateral, superior medial, and posterior lateral protocerebra. We found that most of these areas in the protocerebrum are connected with the AL through multiple tracts, suggesting that these areas are sites of convergence for olfactory information. Furthermore, areas such as the superior medial protocerebrum now appear to receive olfactory output both directly from the AL and indirectly from lobes of the MB and the LH, suggesting a degree of functional interaction among these areas. We also analyzed the length and number of fibers in each tract. We compare our results obtained from wild‐type flies with recent results from transgenic strains and discuss how information about odorants is distributed to multiple protocerebral areas. J. Comp. Neurol. 520:4131–4140, 2012. © 2012 Wiley Periodicals, Inc.     

Connections between the deutocerebrum and the protocerebrum,and neuroanatomy of several classes of deutocerebral projection neurons in the brain of male Periplaneta americana

The topography and neuroanatomy of fibers connecting the deutocerebrum to the protocerebrum in the brain of the American cockroach Periplaneta americana were investigated by staining single or multiple deutocerebral neurons with cobalt, Lucifer Yellow, or biocytin. Five tracts are distinguished on the basis of their routes from origins in the antennal lobe to the protocerebral neuropil: the inner antenno-cerebral tract (IACT); antenno-cerebral tracts II, III, and IV (ACT II, III, IV), and the outer antenno-cereral tract (OACT). These tracts are largely composed of the axons of four classes of deutocerebral projection neurons, which have been identified morphologically; the neuronal arborizations in the glomeruli of the antennal lobe and in the protocerebral projection regions have been examined. Projection neurons with processes in the inner antenno-cerebral tract and in the antenno-cerebral tract II each innervate a single glomerulus in the antennal lobe, and both types have terminals in the calyces of the mushroom bodies and in the lateral lobe of the protocerebrum. The axons of pheromone-sensitive projection neurons with dendritic trees in the male-specific macroglomerulus seem to run exclusively in the inner antenno-cerebral tract. Subgroups of these pheromone sensitive neurons differ in relative sensitivity to the two female attractant components as well as in the arborization pattern of their dendrites in the macroglomerulus. The projection neurons of two other classes each innervate many glomeruli in the antennal lobe, those of one class sending their axons into the protocerebrum in the antenno-cerebral tract IV and the other, in the outer antenno-cerebral tract. The neurons of antenno-cerebral tract IV innervate not only the mushroom body calyces and the lateral lobe but also neuropil regions not previously described in the cockroach. Neurons with axons in the outer antenno-cerebral tract have no terminals in the calyces but innervate the lateral lobe and the neuropil surrounding the tract. The morphological findings presented here show that, in addition to the tracts previously documented in the cockroach brain, there are other, presumably olfactory, connections between the deutocerebrum and the protocerebrum. © 1993 Wiley-Liss, Inc.     

Histochemistry of acetylcholinesterase and immunocytochemistry of an acetylcholine receptor-like antigen in the brain of the honeybee

A histochemical staining method for acetylcholinesterase (AChE) and an antiserum raised against nicotinic acetylcholine receptors (AChR) of locust nervous tissue were applied in order to reveal certain candidates of cholinergic pathways in the brain of the honeybee. The AChE staining marked layers in the optic lobes, fibers connecting the two brain hemispheres, and fiber tracts as well as soma clusters within the protocerebrum. The calycal input regions of the mushroom bodies were labelled, whereas the intrinsic Kenyon cells showed no staining. Although the antennal afferents projecting into the dorsal lobe showed strong AChE activity, projections into the antennal lobe showed rather weak staining. Application of the antiserum against the AChR showed immunoreactivity in neuropiles, tracts, somata, and the antennal nerve. The immunoreactivity of the optic lobes coincided with the banding pattern of the AChE staining. A particularly striking overlap of AChR immunoreactivity and AChE staining was found in the lip neuropile of the mushroom bodies, which would suggest a cholinergic input into this neuropile via fibers of the median antennoglomerular tract. Because the antiserum against locust AChR binds in neuropiles displaying AChE activity, we conclude that this antiserum also cross-reacts with the bee's receptor. This interpretation is supported by experiments showing alpha-bungarotoxin (alpha-BTX) binding sites in some areas of strong immunoreactivity.     

Central processing of sex pheromone,host odour,and oviposition deterrent information by interneurons in the antennal lobe of female Spodoptera littoralis (Lepidoptera: Noctuidae)

Physiological and anatomical characteristics of antennal lobe interneurons in female Spodoptera littoralis (Boisd.) were investigated using intracellular recording and staining techniques. Responses of local interneurons and projection neurons to female sex pheromone components, host plant odours, and behaviourally active oviposition deterrents were recorded. We found local interneurons and projection neurons that responded specifically to only one or two of the tested odours, but we also found less specific cells, and neurons that responded to most of the tested odourants. These findings show that there are not only specific olfactory pathways in female moths up to the protocerebral level, but also that integration can begin in the antennal lobe. No correlation was found between the degree of specificity of either local interneurons or projection neurons and their respective morphological characteristics. Specialized and unspecialized local interneurons arborized throughout the antennal lobe. Specialized and unspecialized projection neurons had uniglomerular arborizations in the antennal lobe and sent their axons to the calyces of the mushroom body, and to the lateral horn of the protocerebrum. One specific projection neuron had multiglomerular arborizations and projected only to the lateral horn of the protocerebrum. Projection neurons arborizing in the glomeruli closest to the entrance of the antennal nerve always responded to pheromone components. No other correlations were found between the arborization pattern of projection neurons in the antennal lobe or in the protocerebrum and their response characteristics. The sensitivity of local interneurons and projection neurons was in the same range as that of receptor neurons in olfactory sensilla on the antennae, suggesting a much lower convergence in the central nervous system in females than in the pheromone-processing pathway in males. © 1994 Wiley-Liss, Inc.     

Multimodal efferent and recurrent neurons in the medial lobes of cockroach mushroom bodies

Previous electrophysiological studies of cockroach mushroom bodies demonstrated the sensitivity of efferent neurons to multimodal stimuli. The present account describes the morphology and physiology of several types of efferent neurons with dendrites in the medial lobes. In general, efferent neurons respond to a variety of modalities in a context-specific manner, responding to specific combinations or specific sequences of multimodal stimuli. Efferent neurons that show endogenous activity have dendritic specializations that extend to laminae of Kenyon cell axons equipped with many synaptic vesicles, termed "dark" laminae. Efferent neurons that are active only during stimulation have dendritic specializations that branch mainly among Kenyon cell axons having few vesicles and forming the "pale" laminae. A new category of "recurrent" efferent neuron has been identified that provides feedback or feedforward connections between different parts of the mushroom body. Some of these neurons are immunopositive to antibodies raised against the inhibitory transmitter gamma-aminobutyric acid. Feedback pathways to the calyces arise from satellite neuropils adjacent to the medial lobes, which receive axon collaterals of efferent neurons. Efferent neurons are uniquely identifiable. Each morphological type occurs at the same location in the mushroom bodies of different individuals. Medial lobe efferent neurons terminate in the lateral protocerebrum among the endings of antennal lobe projection neurons. It is suggested that information about the sensory context of olfactory (or other) stimuli is relayed by efferent neurons to the lateral protocerebrum where it is integrated with information about odors relayed by antennal lobe projection neurons.     

Inventory and distribution of synapses of identified uniglomerular projection neurons in the antennal lobe of Periplaneta americana.

Uniglomerular projection neurons in the antennal lobe of Periplaneta americana, the axons of which connect the lobe to the protocerebrum, were labeled by intracellular injection of Lucifer Yellow or biocytin. The fine structure of individual neurons within the antennal lobe was examined after the injected substances had been converted (by immunohistochemical or histochemical treatment) to electron microscopically visible reaction products. Seven projection neurons were investigated, including attractant neurons, with dendritic arbors in the macroglomerulus, and projection neurons of normal-sized glomeruli. From reconstructions of thin serial sections and examination of additional processes present at various places in the arborization regions, the distribution of synapses within the glomeruli was inferred. Although the projection neurons differ from one another in their glomerular arborization patterns, they are very similar in the spatial segregation of their input and output synapses within the arborization. Output synapses are found on the thick part of the fiber near its site of entry into the glomerulus, as well as in regions within the glomerulus where the neuron has begun to ramify into thinner fibers. In the latter regions, the many output synapses are accompanied by occasional input synapses; hence these are regarded as transitional regions. At the terminal arbors only input synapses were found. This suggests that neurons with dense terminal arborizations receive particularly numerous inputs in these regions. The large number of input synapses reflects the high degree of convergence of afferents onto projection neurons previously demonstrated physiologically. However, the presence of numerous output synapses indicates that projection neurons not only transport sensory information into the protocerebrum but are also a major component of the neuronal circuitry within the antennal lobe.     

Taurine-like immunoreactivity in the brain of the honeybee

Taurine (2-aminoethanesulfonic acid) is one of the most abundant free amino acids in the insect central nervous system. We have investigated the distribution of taurine-like immunoreactivity in the brain of the honeybee with an antiserum recognizing fixed taurine. Taurine-like immunoreactivity appeared within neuronal perikarya, neurites, and terminals, whereas glial cells were unlabelled. All photoreceptor cells of the compound eyes and the ocelli were stained. So were the fibers of the anterior superior optic tract, which connects the optic lobes to the mushroom bodies in the median protocerebrum. In the mushroom bodies the majority of intrinsic Kenyon cells showed high levels of taurine-like immunoreactivity. The lateral antennoglomerular tract, which interconnects the mushroom bodies with the antennal lobes, was also intensely stained. In the antennal lobes, strong labelling was observed within a few fibers that invade a set of posterior glomeruli from the posterior margin. Sensory projections from the antennal nerve into the antennal lobes showed only intermediate levels of staining. Sensory projections into the dorsal lobe were devoid of taurine-like immunoreactivity. Labral, mandibular, maxillary, and labial nerves, which innervate the various parts of the feeding apparatus, contain a set of five to eight heavily stained fibers. A comparison of taurine-like immunoreactivity with glutamate- and GABA-like immunoreactivities in the brain of the honeybee indicates that the three amino acids are enriched in distinct neuronal populations.     

Sensory neuron development revealed by taurine immunocytochemistry in the honeybee

The formation of ommatidia in the compound eyes and sensilla on the antennae of the honeybee was followed and the development of their sensory neurons was traced using an antiserum against taurine as a marker. Taurine-like immunoreactivity (Tau-IR) is expressed in sensory neurons of several modalities, namely visual, olfactory, gustatory, and mechanosensory. Staining intensity is very high in the larva and in the first half of the pupal stage and gradually decreases towards the end of metamorphosis. In the photoreceptor cells of the compound eyes, Tau-IR can be detected from the fifth larval instar onwards, prior to differentiation of other components of the ommatidium. Already in the midstage larvae, when the antennal primordia of the adult still lie within the peripodial cavity, a few presumably mechanosensory neurons are labelled in the pedicellus of the developing antenna. The majority of the antennal sensory neurons which are located on the flagellum start to exhibit Tau-IR upon pupation, long before any cuticular specializations such as sensory hairs or plates are detectable. All known types of antennal sensilla were identified and it could be shown that all of them are innervated by Tau-IR sensory neurons. Thus, taurine immunocytochemistry can be applied as a useful label for developing sensory neurons. Functional implications of taurine during development are discussed. © 1995 Wiley-Liss, Inc.     

Efferent cortical connections of multimodal cortex of the superior temporal sulcus in the rhesus monkey.

The cortex of the upper bank of the superior temporal sulcus (STS) in the rhesus monkey contains a region that receives overlapping input from post-Rolandic sensory association areas and is considered multimodal in nature. We have used the fluorescence retrograde tracing technique in order to answer the question of whether multimodal areas of the STS project back to post-Rolandic sensory association areas. Additionally, we have attempted to answer the question of whether the projections from the multimodal areas directed to the parasensory association areas originate from common neurons via axon collaterals or from individual neurons. The results show that multimodal area TPO of the STS projects back to specific unimodal parasensory association areas of the parietal lobe (somatosensory), superior temporal gyrus (auditory), and posterior parahippocampal gyrus (visual). In addition, a substantial number of projections from area TPO are directed to distal parasensory association areas, area PG-Opt in the inferior parietal lobule, areas Ts1 and Ts2 in the rostral superior temporal gyrus, and areas TF and TL in the parahippocampal gyrus. These latter regions are themselves considered to be higher-order association areas. It was also noted that the majority of the projections to these higher-order association areas originate from the middle divisions of area TPO (TPO-2 and TPO-3). These neurons are organized in a significantly overlapping manner. Despite this overlap of the projection neurons, only an occasional double labeled neuron was observed in area TPO. Thus, our observations indicate that the multimodal region of the superior temporal sulcus has reciprocal connections with the unimodal parasensory association cortices subserving somatosensory, auditory and visual modalities, as well as with other post-Rolandic higher-order association areas. These connections from area TPO to post-Rolandic association areas may have a modulating influence on the sensory association input leading to multimodal areas in the superior temporal sulcus.     

Complete mapping of glomeruli based on sensory nerve branching pattern in the primary olfactory center of the cockroach Periplaneta americana

Glomeruli are structural and functional units in the primary olfactory center in vertebrates and insects. In the cockroach Periplaneta americana, axons of different types of sensory neurons housed in sensilla on antennae form dorsal and ventral antennal nerves and then project to a number of glomeruli. In this study, we identified all antennal lobe (AL) glomeruli based on detailed innervation patterns of sensory tracts in addition to the shape, size, and locations in the cockroach. The number of glomeruli is ∼205, and no sex‐specific difference is observed. Anterograde dye injections into the antennal nerves revealed that axons supplying the AL are divided into 10 sensory tracts (T1–T10). Each of T1–T3 innervates small, oval glomeruli in the anteroventral region of the AL, with sensory afferents invading each glomerulus from multiple directions, whereas each of T4–T10 innervates large glomeruli with various shapes in the posterodorsal region, with a bundle of sensory afferents invading each glomerulus from one direction. The topographic branching patterns of all these tracts are conserved among individuals. Sensory afferents in a sub‐tract of T10 had axon terminals in the dorsal margin of the AL and the protocerebrum, where they form numerous small glomerular structures. Sensory nerve branching pattern should reflect developmental processes to determine spatial arrangement of glomeruli, and thus the complete map of glomeruli based on sensory nerve branching pattern should provide a basis for studying the functional significance of spatial arrangement of glomeruli and its developmental basis. J. Comp. Neurol. 518:3907–3930, 2010. © 2010 Wiley‐Liss, Inc.     

Ramification pattern and ultrastructural characteristics of the serotonin-immunoreactive neuron in the antennal lobe of the moth Manduca sexta: A laser scanning confocal and electron microscopic study

The two antennal lobes, the primary olfactory centers of the brain, of the moth Manduca sexta each contain one neuron that displays serotonin immunoreactivity. The neuron projects out of the antennal lobe and sends branches into ipsi- and contralateral protocerebral areas. An axon-like process extends from the contralateral protocerebrum to, and terminates in, the contralateral antennal lobe. In order to begin to investigate the possible role of this unique neuron in olfactory information processing, we have used laser scanning confocal microscopic and electron microscopic immunocytochemical techniques to study the ramification pattern, ultrastructural characteristics, and synaptic connections of the neuron in the antennal lobes of female adult Manduca sexta. The neuron ramifies extensively in the antennal lobe contralateral to the cell body. The ramifications, mainly in the base and center of each glomerulus, do not overlap with those of the sensory axons from the antenna. This finding suggests that the serotonin-immunoreactive neuron may not receive direct input from sensory neurons, and that it may modulate the activity of the neurons of the antennal lobe rather than that of the sensory neurons. In the electron microscope, the neuron exhibits large dense-cored vesicles and small, clear round vesicles. In the antennal lobe ipsilateral to the cell body, the primary neurite of the serotonin-immunoreactive neuron is unbranched and lacks detectable synaptic connections. The ramifications in the contralateral antennal lobe, however, participate in synaptic connections. At very low frequency, contralateral branches form synapses onto unlabeled processes and also receive synapses from unidentified neurons in the glomeruli, indicating that the neuron may participate directly in synaptic processing of olfactory information. The high ratio of output to input synapses made by the serotonin-immunoreactive processes in the contralateral antennal lobe is consistent with the idea that this neuron may receive synaptic input via its bilateral branches in the protocerebrum and then send information to the contralateral antennal lobe where the neuron may exert feedback or modulatory influences on olfactory information processing in the glomeruli. © 1993 Wiley-Liss, Inc.     

Synaptic organization of the uniglomerular projection neurons of the antennal lobe of the moth Manduca sexta: A laser scanning confocal and electron microscopic study

The detailed branching pattern and synaptic organization of the uniglomerular projection neurons of the antennal lobe, the first processing center of the olfactory pathway, of the moth Manduca sexta were studied with laser scanning confocal microscopy and a technique combining laser scanning confocal microscopy and electron microscopy. Uniglomerular projection neurons, identified electrophysiologically or morphologically, were stained intracellularly with neurobiotin or biocytin. Brains containing the injected neurons were treated with streptavidin-immunogold to label the injected material for electron microscopy and with Cy3-streptavidin to lable the neurons with fluorescence for laser scanning confocal microscopy, and then embedded in Epon. Labeled neurons were imaged and reconstructed with laser scanning confocal microscopy (based on the retained fluorescence of the labeled neuron in the Epon block), and thin sections were cut at selected optical levels for correlation of light microscopic data and electron microscopic detail. Each neuron had a cell body in one of the three cell-body clusters of the antennal lobe, a primary neurite that extended across the coarse neuropil at the center of the antennal lobe and then formed a dense tuft of processes within a single glomerulus, and an axon that emanated from the primary neurite and projected from the antennal lobe via the antenno-cerebral tract to the lateral horn of the ipsilateral protocerebrum and, collaterally, to the calyces of the mushroom body. In the electron microscope, the fine dendritic branches in the apical zones of the glomeruli, where sensory axons terminate, were found to receive many input synapses. In deeper layers across the glomeruli, the processes participated in both input and output synapses, and at the bases of the glomeruli, the most proximal, thickest branches formed output synapses. In both of the protocerebral areas in which axonal branches terminated, those branches formed exclusively output synapses. Our findings indicate that, in addition to conveying olfactory information to the protocerebrum, uniglomerular projection neurons in the antennal lobes of M. sexta participate in local intraglomerular synaptic circuitry. J. Comp. Neurol. 379:2-20, 1997. © 1997 Wiley-Liss, Inc.     

Structural plasticity of identified glomeruli in the antennal lobes of the adult worker honey bee

Adult worker honey bees alter their behaviour with age but retain a strong reliance on sensory information from the antennae. The antennae house a diverse array of receptors, including mechanoreceptors, hygroreceptors, olfactory receptors, and contact chemoreceptors, which relay information to the brain. Antennal sensory neurons that project to the antennal lobes of the brain converge onto second-order interneurones to form discrete spheres of neuropil, called glomeruli. The spatial organisation of glomeruli in the antennal lobes of the honey bee is constant, but the central distribution of information from receptors tuned to different sensory modalities is unknown. Here we show that the glomerular neuropil of the antennal lobes undergoes constant modification during the lifetime of the adult worker bee. Changes in morphology are site specific and highly predictable. The total volume of the glomerular neuropil of the antennal lobe increased significantly during the first 4 days of adult life. Each of the five readily identifiable glomeruli examined in this study exhibited a unique pattern of growth. The growth of two of the five glomeruli changed dramatically with the shift to foraging duties. Furthermore, significant differences were identified between the antennal lobes of bees performing nectar-and pollen-foraging tasks. The highly compartmentalized nature of the antennal lobes, the ease with which specific glomeruli can be identified, and the predictability of changes to the antennal lobe neuropil make this an ideal system for examining the mechanisms and behavioural consequences of structural plasticity in primary sensory centres of the brain. © 1996 Wiley-Liss, Inc.